RE: c-kit

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From:Hewlett Bryan <HEWLETT@HHSC.CA>
To:Histonet <>, "'Kappeler Andreas'" <>

Andi and Heike.

We have been using anti c-kit for the past four years.
For full technical details see paper;
 " ICC as precursors of GIST."  Am. J. Surg. Pathol 23(4): 377-389,1999.
This study used the Santa Cruz antibody. We have also used the Dako rb anti
c-Kit, @ 1:50 with equivalent results.

Andi, FYI I would not use HIER, with either of these ab's. In our hands,
although there is staining of cell bodies, reactivity is reduced and the
long dendritic processes of ICC appear to be destroyed by this procedure.

We have extensively tested tissues(including bone marrow trephines) fixed in
10%NBF, B5 and AZF, both with and without deminerilization in either
Surgipath's Decalcifier II, or 5% formic acid. All work well with the
procedure as described. Stem cells in marrow stain well.

If you have further questions, I will be happy to try and answer.



> ----------
> From: 	Kappeler Andreas[]
> Sent: 	May 25, 2000 6:00 AM
> To: 	Histonet
> Subject: 	Re: c-kit
> Hi Heike
> We have just started to use rb-a-c-kit from Santa Cruz (sc-168) on
> paraffin
> sections of GIST (gastrointestinal stroma tumors), where it works nicely.
> Dilution is 1:50, pretreatment in 10 mM citrate buffer, pH 6.0, pressure
> cooker, with 7 min under full pressure. My first impression with bone
> marrow
> trephines, however, is mixed: we do barely see any c-kit positive stem
> cells, but may be our staining conditions are not (yet) optimized for this
> type of tissue / cells.
> Andi Kappeler
> Institute of Pathology, University of Bern, Switzerland

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